Method and composition for rejuvenating hair, nails, tissues, cells and organs by ex-vivo or immersive treatment

ABSTRACT

A method and composition for the treatment of hair, nail, ex-vivo organ, ex-vivo cell or ex-vivo tissue to improve the biomechanical and diffusional characteristics comprising an effective amount of a compound selected from the group consisting of compounds of the formula (I):

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation, under 35 U.S.C. § 120, of U.S.patent application Ser. No. 10/055,252, filed Jan. 22, 2002 now U.S.Pat. No. 6,713,050, which claims the benefit under 35 U.S.C. § 119(e) ofU.S. Provisional Application Ser. No. 60/263,300, filed Jan. 22, 2001.

BACKGROUND OF THE INVENTION

The present invention generally relates to the aging or damage of hair,nails, tissues, organs and cells and, more particularly, to therejuvenation of hair, nails, tissues, cells, and organs by improving thedeformability and diffusion coefficient of hair, nails, tissues, cells,and organs in humans and animals (including companion animals and livestock).

Various changes in the biomechanical and other functional properties ofhair may occur with aging and diseases. Undesirable changes may includedeterioration in manageability, including decreased stability andbrittle hair. Typically, these detrimental changes may be due to: (a)physically or chemically damaged hair; (b) physiologically aged hair;and/or (c) diseased hair (e.g. hair of diabetics). Hair may bephysically damaged from the normal grooming process of shampooing,combing, drying (e.g. hot air blow drying), and/or brushing. In additionto this physical damage of hair, hair may also be damaged by chemicalaction such as by exposure to sunlight and contact with water containingchemically reactive agents such as oxidizers (e.g bleaching and/ordyeing of hair). Also, the repeated use of permanent waving compositionson the hair fibers may cause damage to the hair especially if not usedaccording to directions. Bleached hair is often characterized as beingdry, brittle, and overly coarse. Finally, with the aging process, hairmay become dry, brittle and overly coarse.

For nails, deterioration in the biomechanical and other functionalproperties may also result in undesirable nail problems. Conventionally,the term “nail” has meant the horny cutaneous plate on the dorsalsurface of the distal end of a finger or toe, or the correspondingappendages in animals. Specifically, in humans, the hardness andstrength of the nails is particularly important not only for the beautyof their appearance, but for the well-being of the individual.Embrittlement of the nails is normally associated with aging. However,various activities also expose nails to a number of materials which mayadversely affect the nail's biomechanical and other functionalproperties. For example, occupational exposure to extensive or constantwetting of the hands with soaps, detergents, solvents, chemical hairwaving and coloring lotions, and insults from deliberate cosmeticapplications, such as manicuring, or any like products can lead todrying, brittleness, cracking, laminating, splitting, ridging andsimilar damage. Additionally, certain diseases may also lead to nailembrittlement or associated disfigurement owing to weakening of nailhardness and strength. Moreover, the appearance of fingernails andtoenails of humans are frequently enhanced with decorative nail-carecosmetics, such as nail polishes, nail polish removers, nail polishbases, alkaline cuticle removers and the like. Overuse of these productscan alter the nail, causing it to weaken, soften, split and break.

With respect to tissues, cells and organs, transplantation of thesematerials has become a routine means of treating certain diseases andother conditions. Transplantation requires a ready source of organs,such as kidney, pancreas, liver, heart, etc., from living persons orcadavers. Conventionally, most vital organs, cells and tissues, whichare used for transplantation, are obtained from heart beating cadaversand preserved for variable periods of time prior to theirtransplantation. However, preservation methods merely attempt tomaintain the present condition of the organ, cell or tissue. For thisreason, the majority of organs, cells and tissues that are used fortransplantation presently come from younger individuals who typicallyhave tissues, cells and organs that have not been detrimentally affectedby age or disease.

In contrast, because of the aging process or disease, older individualshave a deterioration in the biomechanical (e.g. deformability) and otherfunctional properties of their cells, tissues and organs. For thisreason, decreased deformability is associated with impaired tissue ororgan functionality in itself, as optimal biomechanical function isdemonstrated at deformability levels measured in young individuals anddiminishes with progressively decreasing levels of deformability. Thus,at the present time, older individuals typically can not be candidatesfor organ, tissue or cell donation because preservation solutions merelyattempt to preserve the present condition of the organ, cell or tissue.

Conventionally, two typical methods of preserving organs, cells andtissues for transplantation are continuous pulsatile perfusion andsimple hypothermic storage in a preservation solution. In pulsatileperfusion, the organ is subjected to pulsatile flow of a perfusate underhypothermic conditions such that the organ membranes receive sufficientoxygenation. Typically, the perfusate contains albumin and lipids. Withsimple hypothermic storage, organs are removed from a cadaver donor andrapidly cooled. Rapid cooling is achieved by external cooling and byperfusion with a preservation solution to lower the internal temperatureof the organ. The organ is then stored immersed in the preservationsolution at temperatures of about 0°-4° C. Two conventional glucosepreservation flush solutions are the Collins (G. M. Collins, The Lancet,1969, 1219-1222) and the Euro-Collins (J. P. Squifflet et al,Transplant. Proc., 1981, 13:693-696) solutions. These solutions resembleintracellular fluid and contain glucose as an osmotic agent. Despitetheir widespread use, the Collins and Euro-Collins preservationsolutions do not typically provide adequate preservation for storagetimes greater than about 48 hours. For example, kidneys stored inCollins solution for 24 hours may exhibit considerable damage to thenephrons. This damage included degradation of cells lining the proximaltubules, extensive swelling and rupturing of cells lining the ascendingdistal tubules, degeneration of glomerular epithelial and endothelialcells and accumulation of flocculent cytoplasmic debris in the capsularspaces of Bowman. (P. M. Andrews et al, Lab. Invest., 1982, 46:100-120).In addition to glucose flush solutions, high osmolality preservationsolutions have been prepared using raffinose and lactobionate as in theUW preservation solution (R. J. Ploeg et al, Transplant. Proc., 1988, 20(suppl 1) 1:935-938), mannitol in the Sacks solution (S. A. Sacks, TheLancet, 1973, 1:1024-1028), sucrose in the Transplantation, 1989,47:767-771) and the histidine buffered HTK solution of Bretschneider (N.M. Kallerhoff et al, Transplantation, 1985, 39:485-489). Other examplesare solutions that contain synthetic hydroxyethyl starch (HES) as anosmotic colloid.

SUMMARY OF THE INVENTION

In accordance with the present invention, a novel method and compositionare disclosed for the “rejuvenation” of hair, nails, tissues, cells andorgans by ex-vivo treatment. In particular, the composition comprisescompounds for the ex-vivo treatment of hair, nails, tissues, cells andorgans to improve the biomechanical and other functional properties ofhair, nails, tissues, cells and organs. More particularly, for hair andnails, the composition and method of the present invention comprisescompounds for rejuvenating: (a) damaged hair or nails; (b)physiologically aged hair or nails; and/or (c) diseased hair or nails(e.g. diabetes). For tissues, cells and organs, the composition andmethod of the present invention comprises compounds for rejuvenatingtissues, cells and organs by improving the deformability and/ordiffusion coefficient of tissues, cells and organs from a state ofdecreased deformability and impaired diffusional characteristics, astypically observed in cells, tissues or organs of older individuals, toa state of increased deformability and improved diffusionalcharacteristics, as commonly seen in cells, tissues and organs ofhealthy and young individuals (i.e 20 years old).

Decreased deformability is associated with impaired tissue or organfunctionality in itself, as optimal biomechanical function isdemonstrated at deformability levels measured in young individuals anddiminishes progressively with age. It is believed that the agingprocess, in addition to modifying the deformability, also inducesadditional specific impairments in the functionality of cells, tissuesand/or organs that are not directly related to deformability. It isfurther believed that this impairment in functionality is related to analtered diffusion coefficient of molecules across intracellular andextracellular spaces. The altered ability of molecules to traverseintra- and extracellular spaces may affect signaling functions ofhormones and cytokines, transportation of oxygen and nutrients from thevascular space to the cell, and cellular metabolism.

Commonly used methods to determine deformability includeultrasonographic techniques and the measurement of volume-pressure andstress-strain relationships. Commonly used methods to measure thediffusion coefficient of molecules across intracellular andextracellular spaces include the determination of the reaction time ofbiomolecular feed-back mechanisms based on the diffusion of moleculesacross a cellular, tissue and/or organ space to reach a target responseelement and the direct measurement of the diffusion rate of certainmolecules across biological spaces. Examples of methods to determine thebiomolecular feed-back mechanisms include, but are not limited to, thefollowing: (a) hypothalamic-pituitary axis: growth hormone, ACTH, TSH,or prolactin; (b) pituitary-adrenal axis: cortisol; (c)pituitary-thyroid axis: thyroxin; and (d) pituitary-gonadal axis: sexhormones, LH, and FSH. Examples of methods to determine the diffusionrate of certain molecules across biological spaces include, but are notlimited to, the following: arterio-alveolar oxygen and carbon dioxidegradients; insulin resistance; and arterio-venous oxygen gradient (e.g.,heart, muscle).

In particular, the compositions comprise compounds for the ex-vivotreatment of hair, nails, organs, cells and tissues to rejuvenate themby changing deformability and increase the tissue diffusion coefficient.This treatment is accomplished by bathing or perfusing the biologicalmaterial outside of the body. The compounds are members of the class ofcompounds known as thiazoliums.

In one embodiment, the compositions comprise thiazolium compounds havingthe following structural formula (I):

wherein R¹ and R² are independently selected from the group consistingof hydrogen, hydroxy(lower alkyl), lower acyloxy(lower alkyl), loweralkyl, lower alkenyl, or R¹ and R² together with their ring carbons maybe an aromatic fused, ring, optionally substituted by one or more amino,halo or alkylenedioxy groups;

-   Z is hydrogen or an amino group;-   Y is amino, a group of the formula —CH₂C(═O)—R wherein R is a lower    alkyl, alkoxy, hydroxy, amino or aryl group; said aryl group    optionally substituted by one or more lower alkyl, lower alkoxy,    halo, dialkylamino, hydroxy, nitro or alkylenedioxy groups;-   or a group of the formula —CH₂R′-   wherein R′ is hydrogen, or a lower alkyl, lower alkynyl, or aryl    group;-   or a group of the formula —CH₂C(═O)—N(R″)R′″-   wherein (a) R″ is hydrogen and R′″ is a lower alkyl group,    optionally substituted by a C₆-C₁₀ aryl group, or a C₆-C₁₀ aryl    group, said aryl groups optionally substituted by one or more lower    alkyl, halo, or (lower alkoxyl)carbonyl groups; or (b) R″ and R′″    are both lower alkyl groups;-   X is a halide, tosylate, methanesulfonate, mesitylenesulfonate ion,    or other pharmaceutically acceptable anion and mixtures thereof, and    a carrier therefor.

In another embodiment, the composition comprises compounds having thefollowing structural formula (II):

wherein R¹ and R² are independently selected from the group consistingof hydrogen and an alkyl group optionally substituted by a hydroxygroup;

-   Y is a group of the formula —CH₂C(═O)R wherein R is a heterocyclic    group other than alkylenedioxyaryl containing 4-10 ring members and    1-3 heteroatoms selected from the group consisting of oxygen,    nitrogen and sulfur; said heterocyclic group optionally substituted    by one or more substituents selected from the group consisting of    alkyl, oxo, alkoxycarbonylalkyl, aryl, and aralkyl groups; and said    one or more substituents optionally substituted by one or more alkyl    or alkoxy groups;-   or a group of the formula —CH₂C(═O)—NHR′ wherein R′ is a    heterocyclic group containing 4-10 ring members and 1-3 heteroatoms    selected from the group consisting of oxygen, nitrogen, and sulfur;    said heterocyclic group optionally substituted by one or more    alkoxycarbonylalkyl groups;-   and X is a halide, tosylate, methanesulfonate or mesitylenesulfonate    ion, or other pharmaceutically acceptable anion.

The present invention also relates to a method for treating hair andnails and for the ex-vivo rejuvenation of organs, cells and tissues byorgans, cells or tissues with a sufficient amount of one or more of thecompounds of the present invention, or a composition containing asufficient amount to achieve the desired result.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows that a composition of the present invention,3-(2-phenyl-2-oxoethyl)-4,5-dimethyltbiazolium chloride, decreases thestiffness of hair after treatment.

FIG. 2 shows that 3-(2-phenyl-2-oxoethyl)-4,5-dimethylthiazoliumchloride, a composition of the present invention, improved thedistensibility and tensile strength of a tendon after treatment.

DETAILED DESCRIPTION OF THE INVENTION

A method and composition are disclosed for the “rejuvenation” of hair,nails, tissues, cells and organs by ex-vivo treatment. In particular,the composition comprises compounds for ex-vivo treating hair, nails,tissues, cells and organs to improve the biomechanical and otherfunctional properties of hair, nails, tissues, cells and organs. Moreparticularly, for hair and nails, the composition and method of thepresent invention comprises compounds for rejuvenating: (a) damaged hairor nails; (b) physiologically aged hair or nails; and/or (c) diseasedhair or nails (e.g. diabetes). For tissues, cells and organs, thecomposition and method of the present invention comprise compounds forrejuvenating tissues, cells and organs by improving the deformabilityand/or diffusion coefficient of tissues, cells and organs from a stateof decreased deformability and impaired diffusional characteristics, astypically observed in cells, tissues or organs of older individuals, toa state of increased deformability and improved diffusionalcharacteristics, as commonly seen in cells, tissues and organs ofhealthy and young individuals (i.e 20 years old).

In one embodiment, the invention relates to a composition for theex-vivo treatment of hair, nails, organs, cells and tissues to improvethe biomechanical and functional properties comprising one or moreagents comprising of thiazole compounds having the structural formula(I):

wherein R¹ and R² are independently selected from the group consistingof hydrogen, hydroxy(lower alkyl), lower acyloxy(lower alkyl), loweralkyl, lower alkenyl, or R¹ and R² together with their ring carbons maybe an aromatic fused, ring, optionally substituted by one or more amino,halo or alkylenedioxy groups;

-   Z is hydrogen or an amino group;-   Y is amino, a group of the formula —CH₂C(═O)—R-   wherein R is a lower alkyl, alkoxy, hydroxy, amino or aryl group;    said aryl group optionally substituted by one or more lower alkyl,    lower alkoxy, halo, dialkylamino, hydroxy, nitro or alkylenedioxy    groups;-   or a group of the formula —CH₂ R′ wherein R′ is hydrogen, or a lower    alkyl, lower alkynyl, or aryl group;-   or a group of the formula —CH₂C(═O)—N(R″)R′″ wherein (a) R″ is    hydrogen and R′″ is a lower alkyl group, optionally substituted by a    C₆-C₁₀ aryl group, or a C₆-C₁₀ aryl group, said aryl groups    optionally substituted by one or more lower alkyl, halo, or (lower    alkoxy)carbonyl groups; or (b) R′ and R′″ are both lower alkyl    groups;-   X is a halide, tosylate, methanesulfonate, mesitylenesulfonate, or    other pharmacologically acceptable anion and mixtures thereof, and a    carrier therefor.

The lower alkyl groups referred to above contain 1-6 carbon atoms andinclude methyl, ethyl, propyl, butyl, pentyl, hexyl, and thecorresponding branched-chain isomers thereof. The lower alkenyl groupsand lower alkynyl groups independently contain from 2 to 6 carbon atoms.Similarly, the lower alkoxy groups contain from 1 to 6 carbon atoms, andinclude methoxy, ethoxy, propoxy, butoxy, pentoxy, and hexoxy, and thecorresponding branched-chain isomers thereof. These groups areoptionally substituted by one or more halo, hydroxy, amino or loweralkylamino groups.

The lower acyloxy(lower alkyl) groups encompassed by the above formulainclude those wherein the acyloxy portion contains from 2 to 6 carbonatoms and the lower alkyl portion contains from 1 to 6 carbon atoms.Typical acyloxy portions are those such as acetoxy or ethanoyloxy,propanoyloxy, butanoyloxy, pentanoyloxy, hexanoyloxy, and thecorresponding branched chain isomers thereof. Typical lower alkylportions are as described hereinabove. The aryl groups encompassed bythe above formula are those containing 6-10 carbon atoms, such as phenyland lower alkyl substituted-phenyl, e.g., tolyl and xylyl, and areoptionally substituted by 1-2 halo, hydroxy, lower alkoxy or di(loweralkyl)amino groups. Preferred aryl groups are phenyl, methoxyphenyl and4-bromophenyl groups. The halo atoms in the above formula may be fluoro,chloro, bromo or iodo.

For the purposes of this invention, the compounds of formula (I) areformed as biologically and pharmaceutically acceptable salts. Usefulsalt forms are the halides, particularly the bromide and chloride,tosylate, methanesulfonate, and mesitylenesulfonate salts. Other relatedsalts can be formed using similarly non-toxic, and biologically andpharmaceutically acceptable anions.

Representative compounds of the present invention are:

-   3-aminothiazolium mesitylenesulfonate;-   3-amino-4,5-dimethylaminothiazolium mesitylenesulfonate;-   2,3-diaminothiazolium mesitylenesulfonate;-   3-(2-methoxy-2-oxoethyl)thiazolium bromide;-   3-(2-methoxy-2-oxoethyl)-4,5-dimethylthiazolium bromide;-   3-(2-methoxy-2-oxoethyl)-4-methylthiazolium bromide;-   3-(2-phenyl-2-oxoethyl)-4-methylthiazolium bromide;-   3-(2-phenyl-2-oxoethyl)-4,5-dimethylthiazolium bromide;-   3-(2-phenyl-2-oxoethyl)-4,5-dimethylthiazolium chloride;-   3-amino-4-methylthiazolium mesitylenesulfonate;-   3-(2-methoxy-2-oxoethyl)-5-methylthiazolium bromide;-   3-(3-(2-phenyl-2-oxoethyl)-5-methylthiazolium bromide;-   3-[2-(4-bromophenyl)-2-oxoethyl]thiazolium bromide;-   3-[2-(4-bromophenyl)-2-oxoethyl]-4-methylthiazolium bromide;-   3-[2-(4-bromophenyl)-2-oxoethyl]-5-methylthiazolium bromide;-   3-[2-(4-bromophenyl)-2-oxoethyl]-4,5-dimethylthiazolium bromide;-   3-(2-methoxy-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide;-   3-(2-phenyl-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide;-   3-[2-(4-bromophenyl)-2-oxoethyl]-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide;-   3,4-dimethyl-5-(2-hydroxyethyl)thiazolium iodide;-   3-ethyl-5-(2-hydroxyethyl)-4-methylthiazolium bromide;-   3-benzyl-5-(2-hydroxyethyl)-4-methylthiazolium chloride;-   3-(2-methoxy-2-oxoethyl)benzothiazolium bromide;-   3-(2-phenyl-2-oxoethyl)benzothiazolium bromide;-   3-[2-(4′-bromophenyl)-2-oxoethyl]benzothiazolium bromide;-   3-(carboxymethyl)benzothiazolium bromide;-   2,3-diaminobenzothiazolium mesitylenesulfonate;-   3-(2-amino-2-oxoethyl)thiazolium bromide;-   3-(2-amino-2-oxoethyl)-4-methylthiazolium bromide;-   3-(2-amino-2-oxoethyl)-5-methylthiazolium bromide;-   3-(2-amino-2-oxoethyl)4,5-dimethylthiazolium bromide;-   3-(2-amino-2-oxoethyl)benzothiazolium bromide;-   3-(2-amino-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide;-   3-amino-5-(2-hydroxyethyl)-4-methylthiazolium mesitylenesulfonate;-   3-(2-methyl-2-oxoethyl)thiazolium chloride;-   3-amino-4-methyl-5-(2-acetoxyethyl)thiazolium mesitylenesulfonate;-   3-(2-phenyl-2-oxoethyl)thiazolium bromide;-   3-(2-methoxy-2-oxoethyl)-4-methyl-5-(2-acetoxyethyl)thiazolium    bromide;-   3-(2-amino-2-oxoethyl)-4-methyl-5-(2-acetoxyethyl)thiazolium    bromide;-   2-amino-3-(2-methoxy-2-oxoethyl)thiazolium bromide;-   2-amino-3-(2-methoxy-2-oxoethyl)benzothiazolium bromide;-   2-amino-3-(2-amino-2-oxoethyl)thiazolium bromide;-   2-amino-3-(2-amino-2-oxoethyl)benzothiazolium bromide;-   3-[2-(4-methoxyphenyl)-2-oxoethyl]thiazolium bromide;-   3-[2-(2,4-dimethoxyphenyl)-2-oxoethyl]thiazolium bromide;-   3-[2-(4-fluorophenyl)-2-oxoethyl]thiazolium bromide;-   3-[2-(2,4-difluorophenyl)-2-oxoethyl]thiazolium bromide;-   3-[2-(4-diethylaminophenyl)-2-oxoethyl]thiazolium bromide;-   3-propargylthiazolium bromide;-   3-propargyl-4-methylthiazolium bromide;-   3-propargyl-5-methylthiazolium bromide;-   3-propargyl-4,5-dimethylthiazolium bromide;-   3-propargyl-4-methyl-5-(2-hydroxyethyl)thiazolium bromide.-   (2-(3-methoxyphenyl)-2-oxoethyl)thiazolium bromide;-   3-(2-(3-methoxyphenyl)-2-oxoethyl)-4    methyl-5-(2-hydroxyethyl)thiazolium bromide;-   3-(2-(3-methoxyphenyl)-2-oxoethyl)benzothiazolium bromide;-   2,3-diamino-4-chlorobenzothiazolium mesitylenesulfonate;-   2,3-diamino-4-methylthiazolium mesitylenesulfonate;-   3-amino-4-methyl-5-vinylthiazolium mesitylenesulfonate;-   2,3-diamino-6-chlorobenzothiazolium mesitylenesulfonate;-   2,6-diaminobenzothiazole dihydrochloride;-   2,6-diamino-3-(2-(4-methoxyphenyl)-2-oxoethyl)benzothiazolium    bromide;-   2,6-diamino-3-(2-(3-methoxyphenyl)-2-oxoethyl)benzothiazolium    bromide;-   2,6-diamino-3-(2-(4-diethylaminophenyl)-2-oxoethyl)benzothiazolium    bromide;-   2,6-diamino-3-(2-(4-bromophenyl)-2-oxoethyl)benzothiazolium bromide;-   2,6-diamino-3-(2-(2-phenyl-2-oxoethyl)benzothiazolium bromide;-   2,6-diamino-3-(2-(4-fluorophenyl-2-oxoethyl)benzothiazolium bromide;-   3-acetamido-4-methyl-5-(2-acetoxyethyl)thiazolium    mesitylenesulfonate;-   2,3-diamino-5-methylthiazolium mesitylenesulfonate;-   3-(2-(2-naphthyl)-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide;-   3 -(2-(3,5-di-tert-butyl-4-hydroxyphenyl)-2 -oxoethyl)-4-methyl-5    -(2-hydroxyethyl)thiazolium bromide;-   3-(2-(2-(2,6-dichlorophenyl)ethylamino)-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide;-   3-(2-dibutylamino-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)-thiazolium    bromide;-   3-(2-(4-(ethoxycarbonyl)anilino)-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide;-   3-(2-(2,6-diisopropylanilino)-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide;-   3-amino-4-methyl-5-(2-(2,6-dichlorobenzyloxy)ethyl)thioazolium    mesitylenesulfonate;-   3-(2-(4-carbomethoxy-3-hydroxyanilino)-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide;-   2,3-diamino-4,5-dimethylthiazolium mesitylene sulfonate;-   2,3-diamino-4-methyl-5-(2-hydroxyethyl)thiazolium mesitylene    sulfonate;-   2,3-diamino-5-(3,4-trimethylenedioxy phenyl)thiazolium mesitylene    sulfonate;-   3-(2-(1,4-benzodioxan-6-yl)-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide;-   3-(2-(3,4-trimethylenedioxyphenyl)-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide;-   3-(2-(1,4-benzodioxan-6-yl)-2-oxoethyl)thiazolium bromide;-   3-(2-(3,4-trimethylenedioxyphenyl)-2-oxoethyl)thiazolium bromide;-   3-(2-(3,5-di-tert-butyl-4-hydroxyphenyl)-2-oxoethyl)-thiazolium    bromide;-   3-(2-(3,5-di-tert-butyl-4-hydroxyphenyl)-2-oxoethyl)-4-methylthiazolium    bromide;-   3-(2-(3,5-di-tert-butyl-4-hydroxyphenyl)-2-oxoethyl)-5-methylthiazolium    bromide;-   3-(2-(3,5-di-tert-butyl-4-hydroxyphenyl)-2-oxoethyl)-4,5-dimethylthiazolium    bromide;-   3-(2-(3,5-di-tert-butyl-4-hydroxyphenyl)-2-oxoethyl)benzothiazolium    bromide;-   3-(2-(4-n-pentylphenyl)-2-oxoethyl)thiazolium bromide;-   3-(2-(4-n-pentylphenyl)-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide;-   3-(2-4-diethylaminophenyl)-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide;-   3-(2-phenyl-2-oxoethyl)-4-methyl-5-vinylthiazolium bromide;-   3-(2-(3,5-di-tert-butyl-4-hydroxyphenyl)-2-oxoethyl)-4-methyl-5-vinylthiazolium    bromide;-   3-(2-tert-butyl-2-oxoethyl)thiazolium bromide;-   3-(2-tert-butyl-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide;-   3-(3-methoxybenzyl)-4-methyl-5-(2-hydroxyethyl)thiazolium chloride;-   3-(2,6-dichlorobenzyl)-4-methyl-5-(2-hydroxyethyl)thiazolium    chloride;-   3-(2-nitrobenzyl)-4-methyl-5-(2-hydroxyethyl)thiazolium bromide;-   3-(2-(4-chlorophenyl)-2-oxoethyl)thiazolium bromide;-   3-(2-(4-chlorophenyl)-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide; and-   3-(2-(4-methoxyphenyl)-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide.

U.S. Pat. Nos. 5,656,261 and 6,007,865 disclose the compoundsrepresented by Formula I, and their syntheses, which are herebyincorporated by reference herein.

In another embodiment, the compositions comprise thiazolium compoundshaving the following structural formula (II):

wherein R¹ and R² are independently selected from the group consistingof hydrogen and an alkyl group optionally substituted by a hydroxygroup; Y is a group of the formula —CH₂C(═O)R

-   wherein R is a heterocyclic group containing 4-10 ring members and    1-3 heteroatoms selected from the group consisting of oxygen,    nitrogen and sulfur; said heterocyclic group optionally substituted    by one or more substituents selected from the group consisting of    alkyl, oxo, alkoxycarbonylalkyl, aryl, and aralkyl groups; and said    one or more substituents optionally substituted by one or more alkyl    or alkoxy groups;-   or a group of the formula —CH₂C(═O)—NHR′ wherein R′ is a    heterocyclic group containing 4-10 ring members and 1-3 heteroatoms    selected from the group consisting of oxygen, nitrogen, and sulfur;    said heterocyclic group optionally substituted by one or more    alkoxycarbonylalkyl groups;-   and X is a halide, tosylate, methanesulfonate or mesitylenesulfonate    ion or other pharmacologically acceptable anion.

The heterocyclic groups referred to herein include 4-8 membered ringshaving at least one and up to 3 heteroatoms, e.g. oxygen nitrogen, orsulfur, therein, and including various degrees of unsaturation.Representatives of such heterocyclic groups are those such asisoxazolyl, phenylisoxazolyl, furanyl, morpholino, thiomorpholino,pyrimidinyl, piperidino, homopiperidino, piperazino, methylpiperazino,hexamethyleneimino, tetrahydroquinolyl, pyridyl, methylpyridyl,imidazolyl, pyrrolidinyl, 2,6-dimethylmorpholino, 1,2,4-triazoylyl,thiazolyl, thienyl, thiazolinyl, methylthiazolyl, and the like. Theheterocyclic groups of the present invention may be further substituted,for example, by an oxo group, to form, for example, a2-oxo-tetrahydroquinolinyl group, or substituted by one or more alkyl,alkoxycarbonylalkyl, aryl, or aralkyl groups, and such substituents maybe further substituted by one or more alkyl or alkoxy groups.

Examples of Y groups of the compounds of the present invention include,but are not limited to: 3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl];3-(2-(4-morpholinyl)-2-oxoethyl);3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl];3-(2-(1-piperidinyl)-2-oxoethyl);3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl];3-(2-(1-pyrrolidinyl)-2-oxoethyl;3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl];3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl; 3-(2-(2-thienyl)-2-oxoethyl);3-(2-(2-thienyl)-2-oxoethyl); 3-(2-(4-thiomorpholinyl)-2-oxoethyl);3-(2-(hexahydro-1-azepinyl)-2-oxoethyl),3-[2-(4-[2-methoxyphenyl]-1-piperazinyl)-2-oxoethyl;3-(2-(octahydro-1-azocinyl)-2-oxoethyl; 3-(2-(2-pyridinyl)-2-oxoethyl;3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl];3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl;3-[2-(4-benzyl-1-piperidinyl-2-oxoethyl]; and3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl].

The alkyl groups referred to above contain one to about eighteen carbonatoms and include, for example, methyl, ethyl, propyl, butyl, pentyl,hexyl, octyl, decyl, dodecyl, and octadecyl, and the correspondingbranched-chain isomers thereof. Lower alkyl groups, of one to about sixcarbon atoms, are preferred. The alkyl groups optionally substituted byhydroxy groups include alkyl groups as hereinbefore defined substitutedwith a hydroxy group at any position, such as but not limited to thefollowing examples: hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl,1-hydroxypropyl, 2-hydroxypropyl, 3-hydroxypropyl, 6-hydroxyhexyl, andthe like. Hydroxy-substituted lower alkyl groups as defined above arepreferred. Similarly, the alkoxy groups contain from one to abouteighteen carbon atoms, and include, for example, methoxy, ethoxy,propoxy, butoxy, pentoxy, hexoxy, decoxy, and octadecoxy, and thecorresponding branched-chain isomers thereof. Lower alkoxy groups of oneto about 6 carbons, are preferred. The alkoxycarbonylalkyl groupsencompassed by the above formula include those wherein the alkoxyportion contain from one to about eighteen carbon atoms and the alkylportion contains from 1 to about eighteen carbon atoms. Typicalalkoxycarbonyl portions are those such as acetoxy or ethanoyloxy,propanoyloxy, butanoyloxy, pentanoyloxy, hexanoyloxy, decanoyloxy, andoctadecanoyloxy, and the corresponding branched chain isomers thereof.

The aryl groups encompassed by the above formula are those containing6-10 carbon atoms, such as naphthyl, phenyl and alkyl or alkoxysubstituted-phenyl, e.g., toluyl and xylyl.

For the purposes of this invention, the compounds of the presentinvention are formed as biologically and pharmaceutically acceptablesalts. Useful salt forms are the halides, particularly the bromide andchloride, tosylate, methanesulfonate, and mesitylenesulfonate salts.Other related salts can be formed using similarly non-toxic, andbiologically and pharmaceutically acceptable anions. As described in theformula above, the heterocyclic group may be represented by the R groupof the formula —CH₂C(═O)—R, or it may represent the R′ group of theformula —CH₂C(═O)—NHR′.

Representative, non-limiting examples of compounds of the presentinvention are:

-   3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]thiazolium bromide;-   3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4,5-dimethylthiazolium    bromide;-   3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4-methyl-5-(2-hydroxyethyl)thiazolium    bromide;-   3-[2-[4-(2-ethoxy-2-oxoethyl)-2-thiazolyl]amino-2-oxoethyl]-4,5-dimethylthiazolium    chloride;-   3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4-methyl-5-(6-hydroxyethyl)thiazolium    bromide;-   3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl]-4,5-dimethylthiazolium    bromide;-   3-(2-(1-piperidinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide;-   3-(2-(2-furanyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide;-   3-(2-(2-furanyl)-2-oxoethyl)-4-(2-hydroxypentyl)thiazolium bromide;-   3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl]-4,5-dimethylthiazolium    bromide;-   3-(2-(1-pyrrolidinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;-   3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl]-4,5-dimethylthiazolium    bromide;-   3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazolium    bromide;-   3-(2-(2-thienyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;-   3-(2-(2-thienyl)-2-oxoethyl)-4-methyl-5-hydroxyethylthiazolium    bromide 3-(2-(4-thiomorpholinyl)-2-oxoethyl)-4,5-dimethylthiazolium    bromide;-   3-(2-(hexahydro-1-azepinyl)-2-oxoethyl)-4,5-dimethylthiazolium    bromide;-   3-[2-(4-[2-methoxyphenyl]-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazolium    chloride;-   3-(2-(octahydro-1-azocinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium    bromide 3-(2-(2-pyridinyl)-2-oxoethyl)-4,5-dimethylthiazolium    bromide;-   3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazolium    chloride;-   3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazolium    chloride;-   3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazolium    chloride;-   3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazolium    chloride;-   3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4-octylthiazolium bromide;-   3-(2-(4-morpholinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide;-   3-[2-[4-(2-ethoxy-2-oxoethyl)-2-thiazolyl]amino-2-oxoethyl]-4,5-dipropylthiazolium    chloride;-   3-(2-(4-morpholinyl)-2-oxoethyl)-4,5-di(octadecyl)thiazolium    bromide;-   3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl]-4,5-dipentylthiazolium    bromide;-   3-(2-(1-piperidinyl)-2-oxoethyl)-4,5-didodecylthiazolium bromide;-   3-(2-(2-furanyl)-2-oxoethyl)-5-decylthiazolium bromide;-   3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl]-4,5-dioctylthiazolium    bromide;-   3-(2-(1-pyrrolidiny)-2-oxoethyl)-4,5-diethylthiazolium bromide;-   3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl]-4,5-dipentylthiazolium    bromide;-   3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl]-thiazolium bromide;-   3-(2-(2-thienyl)-2-oxoethyl)-thiazolium bromide;-   3-(2-(2-thienyl)-2-oxoethyl)-4-methyl-5-(6-hydroxyhexyl)thiazolium    bromide;-   3-(2-(4-thiomorpholinyl)-2-oxoethyl)thiazolium bromide;-   3-(2-(hexahydro-1-azepinyl)-2-oxoethyl)-4,5-dioctylthiazolium    bromide;-   3-(2-(octahydro-1-azocinyl)-2-oxoethyl)-4,5-didecylthiazolium    bromide;-   3-(2-(2-pyridinyl)-2-oxoethyl)-4,5-dioctylthiazolium bromide;-   3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl]-4,5-dipropylthiazolium    chloride;-   3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl]-4-methylthiazolium    chloride;-   3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]-5-methylthiazolium    chloride;-   3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl]-4-octylthiazolium    chloride;

The compounds represented by Formula II are disclosed in U.S. Pat. No.6,121,300. The syntheses of these compounds, discussed in U.S. Pat. No.6,121,300, are hereby incorporated by reference herein. Pharmaceuticalcompositions may be prepared with a therapeutically effective quantityof the agents or compounds of the present invention and may include apharmaceutically acceptable carrier, selected from known materialsutilized for this purpose. Such compositions may be prepared in avariety of forms, depending on the method of administration. Also,various pharmaceutically acceptable addition salts of the compounds ofthe present invention may be utilized.

In a further embodiment, the present invention relates to methods fortreating hair and nails and ex-vivo treatment of organs, cells andtissues that comprise contacting the targeted area with a composition ofthe present invention. In an embodiment, for topical application to hairor nails, a solution, a lotion or ointment may be formulated one or morewith agents or compounds in a suitable vehicle such as water, ethanol,propylene glycol, perhaps including a carrier to aid in contacting thehair or nails. For example, a topical preparation could include up toabout 10% of the compound of Formula I or II, or mixtures thereof. Othersuitable forms for administration to other body tissues are alsocontemplated.

The agent or compound of Formula I or II is formulated in compositionsin an amount effective to return the biomechanical and diffusionalcharacteristics of the sample to the state of a healthy 20 year old.This amount will, of course, vary with the particular agent beingutilized and the particular dosage form, but typically is in the rangeof 0.01% to 1.0%, by weight, of the particular formulation.

In one embodiment relating to the treatment of hair, the compoundsdiscussed above may be combined with other components to form acomposition that may be used to treat: (a) damaged hair; (b)physiologically aged hair; and (c) diseased hair (e.g. diabetes). Thecompositions of the present invention may be compounded and/or mixedwith shampoo compositions containing, anionic, nonionic and cationicsurfactants, as well as hair conditioning compositions. In yet anotherembodiment, the compositions of the present invention may beincorporated with conventional hair treating compositions such asbleaching compositions, hair dyeing compositions and/or hair relaxers.

In an embodiment that includes a hair treatment composition of thepresent invention, surface active agents may be used and include, butare not limited to, coconut oil fatty acids or oleic acid, alkali metalor ammonium or amine soaps, water-soluble lauryl sulfate salts, usuallyalkali metal, ammonium and ethanolamine, commonly diethanolamine ortriethanolamine, salts; alkanolamine salts of linear C₁₂-C₁₅ alkylbenzene sulfonic acids; water-soluble polyethoxylauryl alcohol sulfatesalts; linear alkyl benzene polyoxyethyl sulfonate salts; sulfatedlauric acid monoglyceride salts; quaternary ammonium compounds such ascetyltrimethyl ammonium chloride; nonionic detergents such asoctylphenoxypoly(ethyleneoxy)ethanol; and amphoteric detergents.

In a further embodiment, a shampoo composition of the present inventionmay include foam boosters or foam stabilizers. Such boosters include,but are not limited to, dialkylolamides of C₈-C₁₈ fatty acids, as, forinstance, lauric or cocodiethanolamides which are represented by theformula R—CO—N—(CH₂—CH₂—OH)₂ where R—CO is a saturated fatty acid acylradical of C₈-C₁₅ fatty acids, particularly lauric acid or myristic acidor mixtures of saturated fatty acids containing predominately from C₁₂to C₁₄ fatty acids and commonly derived from coconut oil.

In one embodiment, the composition for treating hair may be adjusted toa pH from about 6 to about 9, more particularly from about 5.5 to about7.5.

In a further embodiment, the composition for treating hair may includesupplemental ingredients for particular purposes such as polymers,combing aids, etc.—liquids, gels, creams or dry powders.

In an embodiment relating to the treatment of nails, the compoundsdiscussed above may be combined with conventional nail polish componentsto form a composition that may be used to treat: (a) damaged nails; (b)physiologically aged nails; and (c) diseased nails (e.g. diabetes). Thenail polish compositions of the present invention may include varioussolvents, resins, FDA certified pigments, and pigment extenders. Thefollowing are illustrative examples of each of these components. Forexample, solvents may include n-butyl acetate (ester solvent), ethylacetate (ester solvent), propylene glycol methyl ether acetate (estersolvent), isopropyl alcohol (oxygenated solvent), dipropylene glycolmethyl ether (glycol ether solvent), naphtha/petroleum naphtha(aliphatic petroleum solvent) mineral spirits (aliphatic petroleumsolvent). For example, resins may include acrylic resin (a polymer ofacrylic-methacrylic acids and their esters), maleated-rosin(rosin-maleic adduct), nitrocellulose (soluble cellulose ester),pigment. In addition, FD&C colors and/or FDA certified pigments may beused. In a further embodiment, pigment extenders may be used such asmagnesium silicate (Vantac 6H), silicone dioxide (amorphous silica),aluminum stearate, calcium carbonate, barium sulfate, aluminum silicate,calcium silicate and calcium sulfate.

In a further embodiment, the composition of the present invention mayinclude antifungal agents such as miconazole nitrate, ketoconazole,itraconazole, fluconazole, econazole, terconazole, saperconazole,amorolfine, ciclopirox, oxiconazole, clotrimazole, terbinafine,naftifine, and other antifungal drugs that are available in a topicalformulation. In addition, the formulation containing the antifungal drugmay include an agent such as hydroxypropyl-alpha-cyclodextrin thatenhances the water-solubility of the antifungal drug. The anti-fungaldrugs are used in anti-fungally effective amounts. For example,anti-fungally effective amounts may be from about 0.5% to about 10%, byweight, and more particularly from about 1% to about 5%, by weight, ofthe formulation that is applied to the nail or surrounding dermaltissue.

In another embodiment, the present compositions may be applied in thearea of tissue “rejuvenation”. For purposes of the present invention,the terms “cells”, “tissues” and “organs” may be used interchangeably asorgans consist of tissue and tissue contain cells and extracellularmaterial. The term “rejuvenation” means sufficiently improving thedeformability and/or diffusion coefficient of cells, tissues and/ororgans from a state of decreased deformability and impaired diffusionalcharacteristics, as typically observed in cells, tissues or organs ofolder individuals, to a state of increased deformability and improveddiffusional characteristics, as commonly seen in cells, tissues andorgans of healthy and young individuals (e.g., 20 years old). One methodof determining whether the cells, tissues or organs have been“rejuvenated” is whether, after treatment with the compositions of thepresent invention, the biomechanical and diffusional characteristics ofthe treated cells, tissues or organs have been changed by at least 20%toward the characteristics of a healthy 20 year old.

Decreased deformability is associated with impaired tissue or organfunctionality in itself, as optimal biomechanical function isdemonstrated at deformability levels measured in healthy and youngindividuals and diminishes with progressively decreasing levels ofdeformability. It is believed that the aging process, in addition tomodifying the deformability, also induces additional specificimpairments in the functionality of cells, tissues and/or organs thatare not directly related to deformability. It is further believed thatthis impairment in functionality is related to an altered diffusioncoefficient of molecules across intracellular and extracellular spaces.The decreased ability of molecules to traverse intra- and extracellularspaces may affect signaling functions of hormones and cytokines,transportation of oxygen and nutrients from the vascular space to thecell, and cellular metabolism.

Table 1 below is an illustrative list of various types of tissues, cellsand organs that may be treated with the composition and method of thepresent invention. This list is merely illustrative and should not beconstrued as limiting the invention.

TABLE 1 Cells Tissues Organs Beta cells of pancreas Tendons HeartCardiac myocytes Ligaments Lungs Neurons Bone Kidneys MacrophagesVessels Liver Erythrocytes Cardiac valves Spleen Leukocytes CorneaAdrenals Fibrocytes Gonads Skin

For purposes of the present invention, “rejuvenation” may be measured byone or more of the techniques to measure deformability and/or otherfunctionality of tissues, cells or organs. Commonly used methods todetermine deformability include ultrasonographic techniques and thedetermination of volume-pressure and stress-strain relationships, butare not limited to these. Table 2 is an illustrative list of some of thecurrently available measures used for evaluating aspects ofdeformability: This list is merely illustrative and should not beconstrued as limiting the techniques that may be used to measure thedeformability of tissues, cells or organs.

TABLE 2 Cells Tissues Organs In vitro Membrane Compliance Compliancedeformability Distensibility Distensibility Impedance Impedance Tensilestrength Wall tension Wall tension Compressive strength Compressivestrength Flexibility Flexibility Torsion Torsion Elasticity ElasticityViscoelasticity Viscoelasticity Shear Shear In vivo Membrane ComplianceCompliance deformability Distensibility Distensibility ImpedanceImpedance Wall tension Wall tension Flexibility Flexibility ElasticityElasticity Viscoelasticity Viscoelasticity

Another method of determining the degree of “rejuvenation” for thepresent invention is to measure the diffusion coefficient of moleculesacross intracellular and extracellular spaces. Methods used to measurethe diffusion coefficient of molecules across intracellular andextracellular spaces include the determination of the reaction time ofbiomolecular feed-back mechanisms based on the diffusion of moleculesacross a cellular, tissue and/or organ space to reach a target responseelement and the measurement of the diffusion rate of certain moleculesacross biological spaces or the diffusion rate of tracer molecules likedyes or radioisotopes. Examples of methods to determine the biomolecularfeed-back mechanisms include, but are not limited to, the following: (a)hypothalamic-pituitary axis: growth hormone, ACTH, TSH, or prolactin;(b) pituitary-adrenal axis: cortisol; (c) pituitary-thyroid axis:thyroxin; and (d) pituitary-gonadal axis: sex hormones, LH, and FSH.Examples of methods to determine the diffusion rate of certain moleculesacross biological spaces include, but are not limited to, the following:arterio-alveolar oxygen and carbon dioxide gradients; insulinresistance; and arterio-venous oxygen gradient (e.g., heart, muscle).

In one embodiment, the compounds of the present invention may becombined with a pharmacologically acceptable organ storage solution thatresults in a rejuvenation solution. The solution of the presentinvention may be utilized to rejuvenate major organs such as the kidney,heart, pancreas, liver, lungs and intestines and portions or segmentsthereof. In another embodiment, organs may be rejuvenated by flushingthe organ after it has been removed from a cadaver with the rejuvenationsolution of the present invention followed by cold storage of the organin the rejuvenation solution at temperatures of about 4° C. Organsstored in the rejuvenation solution may then be transplanted into anappropriate transplant recipient.

For example, the rejuvenation solution of the present invention may be apharmacologically acceptable solution such as an aqueous buffer solutioncontaining one or more of the specific compounds claimed. In oneembodiment, an aqueous phosphate buffer may be prepared, for example, bymixing sodium hydrogen phosphate (Na₂HPO₄) and sodium dihydrogenphosphate (NaH₂PO₄) in water preferably; the water should be purified bydistillation, deionization, etc. prior to use. In a further example, ifa cardioplegic solution for rejuvenation of hearts is desired, aphosphate buffer solution may be prepared using potassium hydrogenphosphate (K₂HPO₄) and/or potassium dihydrogen phosphate (KH₂PO₄).

In a further embodiment, the rejuvenation composition of the presentinvention may be adjusted to have a pH of 7.0 or greater; moreparticularly in the range of 7.1-7.4.

In another embodiment, the rejuvenating solutions of the presentinvention may contain an osmotic agent. Suitable conventional osmoticagents include any osmotic agent known for use in preservationsolutions, including mannitol, sucrose, raffinose, and lactobionate. Theosmotic agent is added to the rejuvenating solution in a sufficientamount to provide adequate osmolality and rejuvenation properties.

In yet another embodiment, the rejuvenating solution of the presentinvention may contain other components which do not adversely affect therejuvenating properties of the solution. For example, addition ofglutathione or a C₁₋₆alkyl glutathione monoester (M. E. Anderson et al,Arch. Biochem. Biophys., 1985, 239:538-548, ethyl ester) in amounts ofabout 2-10 mmol/l.

In a further embodiment, a suitable hypothermic solution, such as thesolution formulated by the University of Wisconsin, that may be combinedwith the compositions of the present invention to rejuvenate the kidney,is composed of Sodium (30 mmol L⁻¹), Potassium (125 mmol L⁻¹), Magnesium(5 mmol L⁻¹), Sulphate (5 mmol L⁻¹), Lactobionate (100 mmol L⁻¹),Phosphate (25 mmol L⁻¹), Raffinose (30 mmol L⁻¹), Adenosine (5 mmolL⁻¹), Gluthatione (3 mmol L⁻¹), Allopurinol (1 mmol L⁻¹), Insulin (100units L⁻¹), Dexamethasone (8 mg L⁻¹), Bactrim (0.5 ml), HES (50 g L⁻¹),Osmolality (320 mmol kg⁻¹), and pH of 7.4.

Further embodiments of suitable cold storage solutions that may becombined with the compositions of the present invention to rejuvenatethe kidney comprise the Collins solution (Collins, B M, Lancet1969;2:1219) and the Euro-Collins solution (Squifflet J P, TransplantProc 1981;13:693). Examples of further embodiments of suitablepreservation solutions that may be combined with the compositions of thepresent invention to rejuvenate the heart comprise the aforementionedUniversity of Wisconsin solution, the Stanford solution (Stein D G, JThorac Cardiovasc Surg 1991;102:657) and the St. Thomas' solution(Demertzis S, Ann Thorac Surg 1993;55:1131).

Examples of further embodiments of suitable pneumoplegia solutions thatmay be combined with the compositions of the present invention torejuvenate the lung comprise the Euro-Collins and the University ofWisconsin solutions.

In a further embodiment, a suitable preservation solution that may becombined with the compositions of the present invention to rejuvenate aliver is the Euro-Collins solution. In yet a further embodiment, asuitable preservation solution that may be combined with thecompositions of the present invention to rejuvenate a liver consists ofthe University of Wisconsin solution. Examples of further embodiments ofsuitable preservation solutions that may be combined with thecompositions of the present invention to rejuvenate the small bowelscomprise the Euro-Collins and the University of Wisconsin solutions.

Methodology for Determination of Stress vs Strain Relationship(Stiffness)

The distensibility (i.e, elasticity, rigidity, shear, tensile strength,compression, etc.) of hair, nails, tendons, etc. may be determined usingwidely available commercial devices, such as manufactured by Instron.Alternatively, other test methods known to a practitioner of the art maybe employed.

The test assay for determining whether hair has been “rejuvenated” forthe present invention is the following. Individual scalp hairs werecemented into the ends of glass capillary tubes such that severalcentimeters of the shaft protruded from the end. This hair was thentrimmed to extend 18 mm from the end of the tube. The unit was theninserted into a micropipette positioner for study. Evaluation consistedof bending the hair by applying a specified force and displacement.Thus, this determination of stress vs strain of bending assessedelasticity, compressibility, and shear of each hair shaft. Each hairserved as its own control, with baseline and post-treatment propertiesassessed and directly compared.

A 1 mm thick plate with a 0.5 mm diameter hole drilled through wasattached to the surface of an isometric strain gauge. To perform eachmeasurement, the capillary tube was placed over the fenestrated plateand the hair end inserted into the hole in the plate without touchingthe strain gauge. The hair was then straightened, if necessary, suchthat its axis formed a 10 degree angle with respect to vertical. Oncepositioned, the hair was then lowered onto the surface of the straingauge. The force required to bend each hair a specified distance wasdetermined over a wide range of displacements. From the stress vs strainrelationships thus derived, the work to bend each hair under differenttreatment regimens can be directly compared. For purposes of the presentinvention, the term “rejuvenated” hair means an improvement of at least20% in the measured value of the treated hair, toward the values seen inyoung, undamaged hair (i.e. healthy 20 year old).

To determine changes in deformability of nails treated with arejuvenator, a nail specimen is trimmed to standard dimensions(thickness and cross sectional area) and dried for 2 hours in an oven at45° C. The nail is then clamped within an Instron materials testingdevice. A stress vs strain relationship is determined using thisinstrument, taking care to perform testing only within the region ofreversibility (i.e., not to apply excessive stress). The nail specimenis then removed from the materials testing device and exposed to atissue rejuvenator bath at a concentration and for a time needed toprovide the desired change in deformability. The nail specimen is thenwashed in distilled water twice for 10 minutes each and dried in an ovenat 45° C. for 2 hrs. A new stress vs strain relationship is thenobtained after this treatment. This shows that the strain for a specificstress is reduced by about 50%. For purposes of the present invention,the term “rejuvenated” nail means an improvement of at least 20% in themeasured value of the treated nail, toward the values seen in young,undamaged nails (i.e. healthy 20 year old).

The following examples will more fully illustrate the embodiments ofthis invention. Therefore, they should not be construed as limiting ofthe remainder of the disclosure in any way. All parts, percentage andproportions referred to herein and in the appended claims are by weightunless otherwise indicated.

EXAMPLE 1

Effect of Rejuvenator on Hair Deformability

Individual hairs were prepared within capillary tubes as describedabove. The stress versus strain relationship was determined as abaseline. Subsequently, the hair was immersed in a solution of3-(2-phenyl-2-oxoethyl)-4,5-dimethylthiazolium chloride (1 mM) for 15minutes. The hair was then removed from the bath and rinsed for 5minutes in distilled water and dried. The results are shown in FIG. 1,which illustrate that after treatment with a rejuvenator, the hairpossessed increased distensibility, as indicated by the fact that forstress greater than 0.08, the resulting strain is about 50% less thanthat before treatment.

EXAMPLE 2

Rejuvenator Shampoo Composition

The following shampoo composition is prepared employing a compound ofthe present invention and is applied to hair.

Component Weight (%) Sodium Lauryl Sulfate (30%) 40.00 LaurieDiethanolamide 4.00 3-(2-phenyl-2-oxoethyl)-4,5-dimethylthiazolium 1.10chloride Perfume 0.25 Dowicil 200 0.20 Soft Water 54.45

EXAMPLE 3

Rejuvenator Nail Polish Composition

The following nail polish composition is manufactured according toconventional methods containing the compounds of the present invention.

Amount Weight (%) Nitrocellulose (¼″ RS) 15.00 Nitrocellulose (½″ RS)5.00 Dipropylene Glycol Di-P-Aminobenzoate 10.00 Neopentyl GlycolDioctanoate 5.00 Toluene 47.80 Ethyl Acetate 10.00N-Methyl-2-Pyrrolidone 2.00 Isopropanol 0.503-(2-phenyl-2-oxoethyl)-4,5-dimethylthiazolium 2.00 chlorideStearalkonium Hectorite 1.00 Titanium Dioxide 0.30 Black Iron Oxide 0.30D&C Red #7 Ca Lake 0.30 D&C Red #34 Ca Lake 0.30 D&C Yellow #5 Zr Lake0.50

EXAMPLE 4

Rejuvenator Nail Composition with Antifungal

The components outlined below are mixed into a homogeneous solutionwhich is useful for applying to finger and toe nails for rejuvenation.

Component Weight (%) Alcohol SDA 67.0 Miconazole 20.03-(2-phenyl-2-oxoethyl)-4,5-dimethylthiazolium chloride 2.0 Glycerin 1.0Water 4.0 Propylene glycol 5.0 Hydroxypropyl cellulose 1.0

EXAMPLE 5

Rejuvenator Hair Conditioning Composition

Ingredient Weight (%) Deionized Water 93.10 Hydroxyethylcellulose 1.35Silicone Copolymer (SF 1188 (GE)) 0.75 Oleyl Alcohol 0.75 DTAC (LaurylTrimethyl Ammonium Chloride) 2.253-(2-phenyl-2-oxoethyl)-4,5-dimethylthiazolium chloride 1.0 Germaben II(preservative) 0.50 Perfume 0.30

EXAMPLE 6

Tendon Rejuvenation

The elasticity of the Achilles tendon of a one year old hen wasdetermined in a materials testing device before and after ex vivoexposure to 3-(2-phenyl-2-oxoethyl)-4,5-dimethylthiazolium chloride (1mM) in a bath for 15 minutes. The results are shown in FIG. 2.Post-treatment, distensibility was significantly increased towards thatof a younger chicken. Specifically, for stress greater than 0.04, thetreated tendon exhibits about 50% of the strain of the untreated tendon.

EXAMPLE 7

Tissue Diffusion Coefficient Rejuvenation

The following is an illustration of a methodology that can be used torejuvenate ex vivo and prior to transplantation to a recipient patientthe lungs removed from a donor cadaver aged 21 years or more. Oldersubjects exhibit a decrease in the deformability of pulmonary tissue andin the capacity of gases, including oxygen, to diffuse across alveolarmembranes. Individuals' deformability and diffusion characteristics canbe characterized by using the standard methodology of determination ofthe lung diffusing capacity for carbon monoxide (e.g., Chang et al.,Chest (1992) 102:139-42). The diffusing capacity is measured in thesupine patient using a single breath of carbon monoxide (0.3%) held forten seconds using a CPI 5000 IV (Gould Instruments, Houston, Tex.),corrected for alveolar volume. A young subject (20 years old) will haveapproximately a 40% higher diffusion coefficient for carbon monoxidecompared to older subjects (60 yrs). The ex vivo organ is flushed oncewith a cooled preservation solution containing3-(2-phenyl-2-oxoethyl)-4,5-dimethylthiazolium chloride at aconcentration of 0.01 to 1.0 mM. Testing in the recipient patient,following transplantation of the lungs, would show that bothdeformability and diffusional capacity had improved to a statecomparable to the one observed in the lungs of a healthy and youngindividual (i.e., 20 years).

EXAMPLE 8

Rejuvenation of Organs Composition

A 70 year old male individual and established organ donor is declaredbrain dead 24 hours following a motor vehicle accident during which hesustained serious brain trauma. This individual has no known history ofcardiovascular or pulmonary disease, infection with the hepatitisviruses, HIV or neoplastic processes. His organs are considered fordonation. However, because of his age, donation of the kidneys, heartand lungs would generally not be indicated. The composition and methodof the present invention are employed to rejuvenate the aged organs to astate compatible with the organs of a young individual, exhibitingdeformability and diffusion characteristics of young organs. The regimenapplied consists of the ex vivo perfusion of the extracted organs withan accepted preservation solution containing a sufficient concentrationof the composition of the present invention for a sufficient duration oftime, e.g., 15 minutes, following conventionally established guidelinesfor the preservation of cells, tissues and/or organs ex vivo.Thereafter, depending on need, the organs are continually perfused witha conventional preservation solution which may or may not containadditional concentrations of the compositions of the present invention.

The organ extraction as well as the ex vivo handling and implantation ofthe organ into the recipient are in accordance with establishedalgorithms and procedures. Prior to implantation, the organ may beflushed with a conventional preservation solution. This flushingprocedure permits the transplantation of an organ free of thecompositions of the present invention.

It is believed that the transplanted organs, treated as describedherein, would demonstrate improved functionality, both from abiomechanical and molecular diffusion/transportation perspective.Moreover, the tissue rejuvenation technology would also increase thenumber of transplantable organs significantly. There is a great demandfor transplantable organs which is widely exceeding the available supplyof transplantable organs. Alternatively, the technology would alsoimprove the functionality of currently transplantable organs and improvesurvival and clinical outcome of such organs post transplantation.

1. A method for the ex vivo treatment of hair or nail to improve thebiomechanical and functional properties comprising externally contactingto hair or nail with an effective amount of composition comprising acompound selected from the group consisting of compounds of the formula(I):

wherein R¹ and R² are independently selected from the group consistingof hydrogen, hydroxy(lower alkyl), lower acyloxy(lower alkyl), loweralkyl, lower alkenyl, or R¹ and R² together with their ring carbons maybe an aromatic fused ring, optionally substituted by one or more amino,halo or alkylenedioxy groups; Z is hydrogen or an amino group; Y isamino, or a group of the formula —CH₂C(═O)—R wherein R is a lower alkyl,alkoxy, hydroxy, amino or aryl group; said aryl group optionallysubstituted by one or more lower alkyl, lower alkoxy, halo,dialkylamino, hydroxy, nitro or alkylenedioxy groups, or a group of theformula —CH₂R′ wherein R′ is hydrogen, or a lower alkyl, lower alkynyl,or aryl group, or a group of the formula —CH₂C(═O)—N(R″)R′″ wherein (a)R″ is hydrogen and R′″ is a lower alkyl group optionally substituted bya C₆-C₁₀ aryl group, or a C₆-C₁₀ aryl group, said aryl groups optionallysubstituted by one or more lower alkyl, halo, or (lower alkoxy)carbonylgroups; or (b) R″ and R′″ are both lower alkyl groups; and X is ahalide, tosylate, methanesulfonate, mesitylenesulfonate, or otherpharmacologically acceptable anion and mixtures thereof, and a carriertherefore.
 2. A method for the ex-vivo treatment of hair or nail toimprove the biomechanical and functional properties comprisingexternally contacting the hair or nail with an effective amount ofcomposition comprising a compound selected from the group consisting ofcompounds of the formula (II):

wherein R¹ and R² are independently selected from the group consistingof hydrogen and an alkyl group optionally substituted by a hydroxygroup; Y is a group of the formula —CH₂C(═O)R wherein R is aheterocyclic group containing 4-10 ring members and 1-3 heteroatomsselected from the group consisting of oxygen, nitrogen and sulfur; saidheterocyclic group optionally substituted by one or more substituentsselected from the group consisting of alkyl, oxo, alkoxycarbonylalkyl,aryl, aralkyl groups; and said one or more substituents optionallysubstituted by one or more alkyl or alkoxy groups; or a group of theformula —CH₂C(═O)—NHR′ wherein R′ is a heterocycic group containing 4-10ring members and 1-3 heteroatoms selected from the group consisting ofoxygen, nitrogen, and sulfur; said heterocyclic group optionallysubstituted by one or more alkoxycarbonylalkyl groups; and X is ahalide, tosylate, methanesulfonate, or mesitylenesulfonate ion, or otherphaimacologically acceptable anion.
 3. The method of claim 1 wherein thecompound is selected from the group consisting of3-(2-phenyl-2-oxoethyl)thiazolium,3-(2-phenyl-2-oxoethyl)-4-methylthiazolium,3-(2-phenyl-2-oxoethyl)-4,5-dimethylthiazolium,3-(2-amino-2-oxoethyl)-4-methyl-5-(2-hydroxyethyl)thiazolium,2,3-diaminothiazolium, 2,3-diamino-4,5-dimethylthiazolium, or2,3-diamino-4-methyl-5-(2-hydroxyethyl)thiazolium.
 4. The method ofclaim 2 wherein the compound is selected from the group consisting of3-(2-(2-thienyl)-2-oxoethyl)-4,5-dimethylthiazolium,3-(2-(1-pyrrolidinyl)-2oxoethyl)-4,5-dimethylthiazolium, or3-(2-(4-morpholinyl)-2-oxoethyl-4,5-dimethylthiazolium.
 5. The method ofclaim 1 or 2 wherein the effective amount is sufficient to treat atleast one of the following: (a) damaged nails; (b) physiologically agednails; and (c) diseased nails.
 6. The method of claim 1 or 2 wherein theeffective amount is sufficient to treat at least one of the following:(a) damaged hair; (b) physiologically aged hair; or (c) diseased hair.7. The method of claim 1 or 2 wherein the effective amount is sufficientto return the biochemical and diffusional characteristics of the hair ornail to the state of a healthy 20 year old.
 8. The method of claim 1 or2 wherein the effective amount is 0.01% to 2% by weight of thecomposition.
 9. The method of claim 6 wherein the composition is ashampoo or hair conditioning composition.